Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction
Spot real-time RT-PCR is a method for detection of Plum pox virus using conventional ELISA plant crude extracts immobilised on paper without the need of RNA extraction. The method was validated on different prunus field samples collected in winter and springtime. Seventy-nine percent of accuracy was...
| Autores principales: | , , , , , |
|---|---|
| Otros Autores: | |
| Formato: | conferenceObject |
| Lenguaje: | Inglés |
| Publicado: |
2017
|
| Acceso en línea: | http://hdl.handle.net/20.500.11939/4957 |
| _version_ | 1855032202586226688 |
|---|---|
| author | Capote, Nieves Bertolini, Edson Martínez, M. Carmen Olmos, Antonio Gorris, María T. Cambra, Mariano |
| author2 | Ertunc, F. |
| author_browse | Bertolini, Edson Cambra, Mariano Capote, Nieves Ertunc, F. Gorris, María T. Martínez, M. Carmen Olmos, Antonio |
| author_facet | Ertunc, F. Capote, Nieves Bertolini, Edson Martínez, M. Carmen Olmos, Antonio Gorris, María T. Cambra, Mariano |
| author_sort | Capote, Nieves |
| collection | ReDivia |
| description | Spot real-time RT-PCR is a method for detection of Plum pox virus using conventional ELISA plant crude extracts immobilised on paper without the need of RNA extraction. The method was validated on different prunus field samples collected in winter and springtime. Seventy-nine percent of accuracy was obtained on winter samples and 97% in samples collected in spring. The sensitivity of spot real-time RT-PCR technique was compared with those obtained by DASI-ELISA, Co-PCR and conventional real-time RT-PCR using RNA as template. Spot real-time RT-PCR resulted 100-times more sensitive than DASI-ELISA and 10-times more sensitive than Co-PCR, although conventional real-time PCR was the most sensitive method for PPV detection. Joined to these advantages, spot real-time RT-PCR permits the storage of immobilised samples on paper at room temperature or frozen for long periods without decreasing the sensitivity of detection. |
| format | conferenceObject |
| id | ReDivia4957 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia49572025-04-25T14:53:50Z Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction ACTA HORTICULTURAE Capote, Nieves Bertolini, Edson Martínez, M. Carmen Olmos, Antonio Gorris, María T. Cambra, Mariano Ertunc, F. Caglayan, Kadriye Spot real-time RT-PCR is a method for detection of Plum pox virus using conventional ELISA plant crude extracts immobilised on paper without the need of RNA extraction. The method was validated on different prunus field samples collected in winter and springtime. Seventy-nine percent of accuracy was obtained on winter samples and 97% in samples collected in spring. The sensitivity of spot real-time RT-PCR technique was compared with those obtained by DASI-ELISA, Co-PCR and conventional real-time RT-PCR using RNA as template. Spot real-time RT-PCR resulted 100-times more sensitive than DASI-ELISA and 10-times more sensitive than Co-PCR, although conventional real-time PCR was the most sensitive method for PPV detection. Joined to these advantages, spot real-time RT-PCR permits the storage of immobilised samples on paper at room temperature or frozen for long periods without decreasing the sensitivity of detection. 2017-06-01T10:11:26Z 2017-06-01T10:11:26Z 2008 2008 conferenceObject Capote, N., Bertolini, E., Martinez, M.C., Olmos, A., Gorris, M. T., Cambra, M. (2008). Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction. Proceedings of the Twentieth International Symposium on Virus and Virus-Like Diseases of Temperate Fruit Crops - Fruit Tree Diseases, (781), 215-219. 0567-7572; 978-90-6605-080-8 http://hdl.handle.net/20.500.11939/4957 10.17660/ActaHortic.2008.781.32 en openAccess Impreso |
| spellingShingle | Capote, Nieves Bertolini, Edson Martínez, M. Carmen Olmos, Antonio Gorris, María T. Cambra, Mariano Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction |
| title | Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction |
| title_full | Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction |
| title_fullStr | Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction |
| title_full_unstemmed | Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction |
| title_short | Spot real-time RT-PCR: A method for direct detection of Plum pox virus avoiding RNA extraction |
| title_sort | spot real time rt pcr a method for direct detection of plum pox virus avoiding rna extraction |
| url | http://hdl.handle.net/20.500.11939/4957 |
| work_keys_str_mv | AT capotenieves spotrealtimertpcramethodfordirectdetectionofplumpoxvirusavoidingrnaextraction AT bertoliniedson spotrealtimertpcramethodfordirectdetectionofplumpoxvirusavoidingrnaextraction AT martinezmcarmen spotrealtimertpcramethodfordirectdetectionofplumpoxvirusavoidingrnaextraction AT olmosantonio spotrealtimertpcramethodfordirectdetectionofplumpoxvirusavoidingrnaextraction AT gorrismariat spotrealtimertpcramethodfordirectdetectionofplumpoxvirusavoidingrnaextraction AT cambramariano spotrealtimertpcramethodfordirectdetectionofplumpoxvirusavoidingrnaextraction AT capotenieves actahorticulturae AT bertoliniedson actahorticulturae AT martinezmcarmen actahorticulturae AT olmosantonio actahorticulturae AT gorrismariat actahorticulturae AT cambramariano actahorticulturae |